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ADK | Adenylate kinase

AS16 3155   | Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: E. coli

ADK | Adenylate kinase in the group Bacterial/Fungal Antibodies at Agrisera AB (Antibodies for research) (AS16 3155)

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product information
Background

Adenylate kinase (ADK) is important in cellular energy homeostasis. It is a phosphotransferase enzyme, catalyzing the interconversion of adenine nucleotides.

Alternative name: myokinase

Immunogen

Protein derived from Escherichia coli, Uniprot: P69441

Host Rabbit
Clonality Polyclonal
Clone
Purity

Serum

Format Lyophilized
Quantity

50 µl

Reconstitution

For reconstitution add 50 µl, of sterile water.

Storage

Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

Tested applications Western blot (WB)
Related products

Secondary antibodies

Additional information
application information
Recommended dilution 1: 100 000 (WB)
Expected | apparent MW

23.5 kDa (E.coli), 24.3 kDa (S.cerevisiae)

Confirmed reactivity Escherichia coli
Predicted reactivity  
Not reactive in

no confirmed exceptions from predicted reactivity are currently known

Additional information
Selected references Tükenmez et al. (2016). Linkage between Fitness of Yeast Cells and Adenylate Kinase Catalysis. PLoS One. 2016 Sep 19;11(9):e0163115. doi: 10.1371/journal.pone.0163115. eCollection 2016.

Application example

Western blot using anti-ADK antibodies

Protein samples were prepared using TCA-extraction method. S. cerevisiae adk1Δ strains expressing either S. cerevisiae Adk1 protein (24.3 kDa) or E. coli adk protein (23.5 kDa) were cultivated logarithmically, and 5 OD-units of cells were collected for TCA-extraction method. In the end, protein pellets were re-suspended with 300 μl of TCA-Laemmli Loading Buffer and denatured at 100°C for 10 minutes. Total protein samples (8 μl of each sample) were separated by 12% SDS-PAGE and transferred to nitrocellulose membranes using a wet transfer protocol. The membranes were blocked with 5% dry-milk in 1x TBST for 1.5 hours at room temperature (RT) with agitation and then they were incubated with primary antibody (anti-adk antibody, 1:2,000) in 5% dry-milk in 1x TBST overnight at 4°C with agitation. The primary antibody solution was decanted and the membranes were washed 3 times for 10 minutes with 1x TBST with agitation. The membranes were then incubated with secondary antibody (HRP conjugated anti-rabbit IgG, 1:4,000) in 5% dry-milk in 1x TBST for 1 hour at RT with agitation. The membranes were washed as above and the signals were detected using Amersham ECL Western blotting detection reagents (GE-healthcare) by exposing the membranes in LAS4000 machine (GE-healthcare) for 10 seconds. Followed-up experiments revealed that lowering the primary antibody concentration from 1:2,000 to 1:50,000-100,000 reduces the unspecific background signals and lowers the risk of target signal oversaturation.

Courtesy of Dr. Hasan Tukenmez, Umeå University, Sweden

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